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Method for extracting biotin from corn steep liquor and thin layer chromatography (TLC) scanning detection method of biotin

A technology of thin-layer chromatography and corn steep liquor, which is applied in the direction of measuring devices, analysis materials, and preparation of test samples. It can solve the problems of time-consuming operation, large detection error, and laborious operation, and achieve the effect of simple operation.

Inactive Publication Date: 2014-07-16
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Because most of the strains are not specific in microbiological method, it will bring a large detection error, and the whole operation process is time-consuming and labor-intensive

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 extracts biotin from corn steep liquor and detects

[0024] Extraction: Take 100ml of a certain batch of corn steep liquor, add 200ml of N,N-dimethylformamide to the corn steep liquor, stir and extract in a water bath, the temperature is 75°C, the extraction time is 1 hour, and the filtrate is obtained by filtration. And wash the filter residue with absolute ethanol, wash 3 times with 100ml each time, combine the filtrate and washing solution. After the mixture was filtered, it was evaporated and concentrated to 80ml with a rotary evaporator, and 240ml of absolute ethanol was added to stir and precipitate some impurities. Rotary evaporator concentrated, concentrated to 20ml, added 100ml acetone to precipitate impurities, filtered and concentrated to 5ml by rotary evaporator; mixed with 5ml butyl acetate and 5ml cyclohexane, left to separate layers, took the supernatant and concentrated to 2ml (as a sample liquid); the rotary evaporation temperature is 40°C...

Embodiment 2

[0027] Embodiment 2 extracts biotin from corn steep liquor and detects

[0028] Take 100ml of another batch of corn steep liquor, add 150ml of N,N-dimethylformamide to the corn steep liquor, carry out stirring and extraction in a water bath, the temperature is 80°C, the extraction time is 1.5 hours, and the filtrate is obtained by filtration, and anhydrous Wash the filter residue with ethanol, wash 5 times with 100 ml each time, and combine the filtrate and washing liquid. After the mixture was filtered, it was evaporated and concentrated to 60ml with a rotary evaporator, and 180ml of absolute ethanol was added to stir and precipitate some impurities. Rotary evaporator concentrated, concentrated to 20ml, added 80ml acetone to precipitate impurities, filtered and concentrated to 5ml by rotary evaporator; mixed with 5ml butyl acetate and 5ml cyclohexane, left to separate layers, took the supernatant and concentrated to 2ml (as a sample liquid); the rotary evaporation temperatur...

Embodiment 3

[0030] Embodiment 3 extracts biotin from corn steep liquor and detects

[0031]Take another batch of corn steep liquor 100ml, add 100ml N,N-dimethylformamide to the corn steep liquor, carry out stirring and extraction in a water bath, the temperature is 85°C, the extraction time is 2 hours, the filtrate is obtained by filtration, and The filter residue was washed with water and ethanol, 100 ml each time for 4 washes, and the filtrate and washing liquid were combined. After the mixture was filtered, it was evaporated and concentrated to 70ml with a rotary evaporator, and 210ml of absolute ethanol was added to stir and precipitate some impurities. Rotary evaporator concentrated, concentrated to 20ml, added 60ml of acetone to precipitate impurities, filtered and concentrated to 5ml by rotary evaporation; mixed with 5ml butyl acetate and 5ml cyclohexane, left to separate layers, took the supernatant and concentrated to 2ml (as a sample liquid); the rotary evaporation temperature ...

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PUM

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Abstract

The invention discloses a method for extracting biotin from corn steep liquor. The method is characterized by using corn steep liquor as a raw material, carrying out extraction with N,N-dimethylformamide, carrying out extraction and impurity removal with absolute ethyl alcohol, carrying out extraction and impurity removal with acetone and carrying out extraction and impurity removal with butyl acetate and cyclohexane, thus finally obtaining a sample solution containing biotin. The invention also provides a thin layer chromatography (TLC) scanning detection method of biotin. The detection method comprises the following steps: (1) dropping the sample solution to be detected on an efficient thin layer plate coated with silica gel GF254, adding a developer, developing the sample solution to be detected in a developing cylinder, and then taking out the thin layer plate and putting the thin layer plate in a fume hood to be aired; (2) spray-dyeing the developed and aired plate with a dyeing solution to obtain spots, in obvious contrast to the background, of standard substances and samples; (3) carrying out TLC scanning by utilizing a Camag3 type thin layer scanner in the absorption wavelength of 530nm, and computing the mass concentration of biotin in the sample solution. The methods have the advantages of simplicity, quickness, accuracy, reliability, stability and the like.

Description

technical field [0001] The invention relates to a method for extracting biotin from corn steep liquor and a thin-layer chromatography scanning detection method for biotin. Background technique [0002] Biotin is one of the essential vitamins in life activities. It is a cofactor of many enzymes in the body, participates in the carboxylation, decarboxylation and dehydrogenation reactions of the body, and participates in the metabolism of sugar, fat and protein. Biotin has been widely used in food, chemical industry, medicine, animal husbandry, biological fermentation and other fields. In the production of glutamic acid fermentation, the control of the amount of biotin directly affects the growth, proliferation, metabolism, cell wall, cell membrane permeability and the rate of glutamic acid production of the producing bacteria, because most glutamic acid bacteria are Biotin-deficient, so the content of biotin plays a vital role in the production, so it is of great significance...

Claims

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Application Information

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IPC IPC(8): G01N30/90G01N1/28C07D495/04
Inventor 冯木香刘代成
Owner SHANDONG NORMAL UNIV
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